Electronic Supplementary Material (ESI) for Food & Function.
This journal is © The Royal Society of Chemistry 2017
Supplementary information
Table S1 GC-MS assignments of metabolites identified in rat urine via GC/MS post silylation
Peak
Metabolites
KI
RT(min)
m/z
1
U2
U3
U4
U5
U6
U7
Hydorgen sulfide -TMS
Unknown
Silanamine TMS
Unkown
Lysine, 4-hydroxy TMS
Borate-TMS
Nicotinic acid, 4-amino-, methyl ester
TMS
Lactic acid TMS
Glycolic acid-TMS
L-alanine-TMS
N-butylamine TMS
Fumaric acid TMS
Glycine, N TMS
Oxalic acid TMS
Malonic acid TMS
Phenol, 3-(1,1-dimethylethyl)-4-methoxyTMS
Unknown
Lvaline TMS
Urea TMS
Benzoic acid TMS
Oxamic acid TMS
Glycerol TMS
Phosphate TMS
Glycine TMS
Succinic acid TMS
Glyceric acid TMS
Uracil TMS
O-Methyl methylphenylthiophosphinate
TMS
Phenylamine TMS
N-hydroxysuccinimide TMS
Malic acid TMS
Threitol TMS
Erythritol TMS
Aspartic acid TMS
Pyroglutamic acid TMS
Ribose TMS
5-MethylcytosineTMS
Threonic Acid TMS
940
947
955
956
962
972
8.64
8.86
9.06
9.11
9.28
9.58
146
151
174
171
144
221
1032
1047
1064
1093
1102
1111
1113
1121
1132
11.28
11.72
12.21
13.03
13.28
13.58
13.62
13.86
14.19
152
117
147
116
174
245
102
147
133
1151
1201
1207
1237
1248
1251
1261
1262
1299
1306
1317
1331
14.77
16.29
16.45
17.37
17.71
17.77
18.06
18.11
19.23
19.41
19.75
20.15
165
131
144
147
179
147
205
299
174
147
292
241
1354
1362
1411
1475
1483
1490
1508
1517
1522
1533
1543
20.81
21.05
22.44
24.19
24.4
24.62
25.08
25.33
25.45
25.73
25.98
188
224
172
233
217
147
232
156
117
254
292
U8
U9
U10
U11
U12
U13
U14
U15
U16
U17
U18
U19
U20
U21
U22
U23
U24
U25
U26
U27
U28
U29
U30
U31
U32
U33
U34
U35
U36
U37
U38
U39
U40
U41
U42
U43
U44
U45
U46
U47
U48
U49
U50
U51
U52
U53
U54
U55
U56
U57
U58
U59
U60
U61
U62
U63
U64
U65
U66
U67
Creatinine enol TMS
Suberic acid 2 TMS
Citramalic acid TMS
Cadaverine TMS
Fructose TMS
Ribitol TMS
Putrescine TMS
2-Keto-glconic acid-TMS
Arabinofuranose TMS
Citric Acid TMS
Pinitol TMS
Ribonic acid TMS
Fumaric acid TMS
Tagatose methoxyamine TMS
Fructose TMS
Ascorbic Acid TMS
Mannose TMS
Allantoin TMS
Glucose TMS
Glucose TMS
Methyl citric acid TMS
Glucosamine TMS
Inositol TMS
Ribonic acid TMS
Glucose TMS
Myo-Inositol TMS
Uric acid-TMS
Mannose- TMS
Sucrose TMS
1550
1574
1591
1622
1643
1687
1733
1761
1792
1813
1829
1839
1851
1861
1867
1871
1874
1878
1899
1906
1922
1939
1949
1951
2077
2083
2129
2340
26.17
26.79
27.23
28.01
28.55
29.61
30.69
31.37
32.09
32.31
32.57
32.98
33.14
33.4
33.61
33.75
33.85
33.9
33.99
34.44
34.61
34.95
35.29
35.51
35.57
38.12
38.25
39.70
47.56
115
303
247
174
103
217
174
292
217
273
260
292
245
307
307
274
205
331
319
319
287
319
318
333
204
318
456
204
361
Figure S1 Base peak chromatogram (BPC) for water extract of grapefruit (A), pomegranate (B)
and red cabbage (C) analyzed by UPLC-qTOF-(-)ESI-MS showing the most significant peaks.
Peak numbers follow those listed in Table S2 for metabolite identification using UPLC-PDAMS.
Figure S2 Diagram sketch representing experimental design and results showing chemical
biomarkers found elevated in rat obese urine i.e, phosphate and oxalate versus i.e., lactate in
healthy ones as revealed from analysis of rat urine samples using GC-MS coupled to OPLS-DA
Table S2 Metabolites identified in pomegranate, red cabbage, grapefruit juice extracts via UPLC–PDA–MS in negative ionization mode.
Peak
UV(nm)
[M-H]-
Mol.
Formula
Erro
r
(pm)
MS fragments
25.6
29
265.5
270, 325 shd.
341.108
191.02
C12H21O11
C6H7O7
2.9
1.2
179
133, 111
71.8
258, 290 shd.
353.145
C14H25O10
7
191
102.6
273.5
299.077
C13H15O8
8.1
279, 137
131.8
146.9
287, 326
292, 325
375.0689
789.2104
C25H11O4
C33H41O22
7.1
1.1
149
289, 322
771.201
C33H39O21
-2.7
8.
9.
10.
11.
12.
153.5
278, 323 shd.
341.087
C15H17O9
2.1
353, 275, 203
415, 353, 191
695, 547, 415,
375, 353, 285,
191
299, 179, 161
160.8
160.9
168
289, 328
299, 330
277
789.2112
341.0892
359.0976
C33H41O22
C15H17O9
C15H19O10
2.1
4.1
2.2
179.2
275
783.0712
C34H23O22
3.3
13.
14.
15.
16.
187.6
192.7
276
275, 301 shd.
323.1333
363.1071
C13H23O9
C18H19O8
4.5
4
389, 315
263, 179, 161
315, 197
631, 621, 481,
470
281
145
196.6
288
353.0872
C16H17O9
1.8
291, 191, 161
200.3
282, 314 shd.
977.2594
C44H49O25
-2.6
771, 577, 385,
285, 223
208.2
279
481.098
C21H21O13
1.6
210.2
295, 330
355.1025
C16H19O9
2.6
1.
2.
3.
4.
5.
6.
7.
RT
(sec.)
17.
18.
Mol.
Erro
465, 423, 387,
335, 325, 265,
207
325, 221
Name
Class
Red
cabbage
pomegran
ate
Grapefruit
Unknown sugar
Sugar
-
-
+
(iso)citric acid
Propanol-O-pentosylhexoside
Hydroxybenzoyl
hexoside
Unknown
Unknown
Kaempferol-Osophoroside -Oglucoside
O-Caffeoyl-hexoside
Unknown
O-Caffeoyl-hexoside
O-Syringoyl-hexoside
Organic acid
Alcohol
glycoside
+
+
-
-
+
-
Aromatic
-
+
-
+
+
-
-
+
-
-
Phenolic acid
Phenolic acid
+
-
+
+
+
-
Tannin
-
+
-
+
+
+
-
-
+
-
-
-
+
-
-
-
+
Red
Pomegran
Granatin A
Flavonol
acylated
glycoside
Phenolic acid
Unknown sugar
Unknown lignin
O-Caffeoylquinic acid
Kaempferol -Osinapoylglucoside-Osophoroside
Phenolic acid
Flavonol
acylated
glycoside
Granatum flavanyl
xyloside
Flavanone
glycoside
Unknown
Peak
19.
RT
(sec.)
UV(nm)
[M-H]-
Formula
r
(ppm
)
MS fragments
211.5
280, 323 shd.
393.1754
C17H29O10
3
372, 355, 99
212.2
276
785.086
C34H25O22
2.1
633, 483, 481,
321
213.5
289, 329
519.1685
C22H31O14
6.7
216.9
272
633.0724
C27H21O18
1.4
217.5
284, 324 shd.
1109.304
C49H57O29
-4
223.4
281
449.1085
C21H21O11
1
20.
21.
223.7
271, 334
593.1509
C27H29O15
0.5
225.4
-
623.1565
C28H31O16
8.4
27.
28.
225.7
279
393.1178
C19H21O9
3
497, 423, 355,
175
331,481, 265,
179
995, 885, 771,
683, 555, 487,
443, 285
415, 385, 329
575, 503, 473,
311
605, 533, 503,
413, 329
329
228.4
254, 289 shd.
463.0515
C20H15O13
0.7
301, 283
29.
30.
238.7
303
935.2496
C35H51O29
2.7
625, 467, 287
240.5
286, 322
741.2241
C33H41O19
3.9
625, 577, 509,
417, 341
244.3
277
415.1605
C19H27O10
1.2
393
246.8
290, 330
965.2617
C54H45O17
4.6
248.3
274.5
951.0788
C41H27O27
4.5
657
799, 771, 649,
507, 415
249.2
289, 324
753.227
C34H41O19
-2.9
609, 223, 175
[M-H]-
Mol.
Formula
Erro
r
MS fragments
22.
23.
24.
25.
26.
31.
32.
33.
34.
Peak
RT
(sec.)
UV(nm)
Name
Class
cabbage
ate
Grapefruit
Hexen-1-ol-pentosyl
hexoside
Di-O-galloylhexahydroxydiphenoylglucopyranose
(Tercatain)
Alcohol
glycoside
-
+
-
Tannin
-
+
-
Citrusin F
Cinnamate
-
-
+
Punicacortein A/B
Tannin
-
+
-
flavonol acylated
glycoside
+
-
-
-
+
-
-
-
+
-
+
-
-
+
-
-
+
-
+
-
-
-
-
+
+
+
-
-
Kaempferol-Oferuloylsophoroside-dihexoside
Unknown
3,8-C,CDiglucosylapigenin
C,CDiglucosyldiosmetin
Unknown
Ellagic acid-Ohexoside
Unknown
Naringenin 4′- O –
glucoside-7-Orutinoside
(Narirutin 4′-glucoside)
Unknown
Unknown
C-Flavone
glycoside.
C-Flavone
glycoside
Tannin
Flavanone
glycoside
Granatin B
Tannin
-
+
-
Disinapoylgentiobiose
Phenolic acid
+
-
-
Name
Class
Red
cabbage
pomegran
ate
Grapefruit
(ppm
)
35.
36.
37.
38.
39.
40.
41.
255.6
322, 288 shd.
917.2371
C42H45O23
-1.5
285, 161
257.7
324
497.1303
C22H25O13
0.5
427, 161
261
322, 289 shd.
947.2511
C43H47O24
-5.1
285, 161
263.1
272.6
325
279, 300 shd.
413.2176
507.1507
C21H33O8
C24H27O12
1.3
0.1
367, 161
489, 345,327
273.3
288, 326
649.2513
C32H41O14
1.8
289.8
284, 328 shd.
579.1749
C 27H31O14
-5.2
413, 341
459, 413, 341,
271, 191
305.7
287, 327
623.1622
C28H31O16
-0.7
605, 533, 503,
579, 443
306.5
313
229.1437
C12H21O4
3.4
211
308.7
294
507.1147
C23H23O13
-0.6
309.9
322
723.2177
C33H39O18
-4.9
315.4
317.2
287, 323 shd.
275
609.1846
409.2058
C46H25O2
C18H33O10
3.3
5.2
463, 305, 201
427, 341, 223,
193
507, 425, 341
279
333
287, 321
693.2775
C34H45O15
-1.6
531, 443, 341
335
290, 320
711.2883
C34H47O16
-1.9
693, 549, 341
340.7
308
651.1583
C29H31O17
-2.5
343.8
294, 318
843.1994
C39H39O21
-0.6
507, 417, 341
651, 507, 341,
201
[M-H]-
Mol.
Formula
42.
43.
44.
45.
46.
47.
48.
49.
50.
51.
Peak
RT
(sec.)
UV(nm)
Erro
r
MS fragments
Kaempferol-O-pcoumaroylsophorosideO-glucoside
Unknown
Kaempferol-Oferuloylsophoroside-Ohexoside
Unknown
Unknown
Unknown
Naringin
6,8-C,CDiglucosyldiosmetin
isomer
Unknown
Unknown flavonoid
glycoside
Feruloylsinapoylgentiobiose
Unknown
Unknown
Nomilinic acid-4-Oglucoside
Nomilinic acid -17-Oglucoside
Kaempferol acetyl
dihexoside
Flavonol
acylated
glycoside
+
-
-
-
-
+
+
-
-
-
+
-
+
+
O-Flavanone
glycoside
-
-
+
C-flavone glyc.
-
-
+
+
-
-
-
+
+
-
-
-
+
+
-
Limonoid
-
-
+
Limonoid
-
-
+
-
-
+
-
-
+
Red
cabbage
pomegran
ate
Grapefruit
Flavonol
acylated
glycoside
Phenolic acid
Unknown
Name
Class
345
286, 324
537.167
C18H33O18
(ppm
)
0.5
348.7
326
959.2868
C45H51O23
-4.3
351.9
291, 320
633.2545
C32H41O13
1.2
356.5
289, 322
929.2749
C44H49O22
-3
56.
57.
58.
360
367.1
268, 314
292, 319
201.0186
399.2375
C 11H 5O4
C21H35O7
3.5
3.4
343, 299
753, 537, 411,
343, 223
463, 341, 261,
193
641, 537, 489,
343, 223
341, 330, 201
373.1
286, 326
593.1872
C28H33O14
0.7
341
59.
60.
61.
403.2
407.9
286
261, 288 shd.
481.1134
269.0434
C25H21O10
C15H9O5
1.3
7.8
-
417.9
N.D.
327.2196
C18H31O5
5.9
291, 309
420.4
301
371.1488
C21H23O6
3.3
341
329.234
329.2312
C18H33O5
C18H33O5
1.8
6.6
311, 293
-
469.186
C26H29O8
1.6
341
307.1895
C18H27O4
6.5
289, 235
311.2214
487.3411
C18H31O4
C30H47O5
4.5
3.8
293, 265
447
52.
53.
54.
55.
62.
63.
64.
65.
N.D.
443.2
444
489.5
66.
N.D.
N.D.
N.D.
501.2
67.
68.
69.
N.D.
552.2
555
N.D.
N.D.
564.4
Peak
RT
(sec.)
309.2063
UV(nm)
[M-H]-
C18H29O4
Mol.
Formula
2.8
291, 197
Erro
r
(ppm
MS fragments
Unknown
Trisinapoylgentiobiose
Obacunone 17-Oglucoside
Feruloyldisinapoylgentiobiose
Bergaptol
Citrusoside A
(Neo)poncirin
Unknown
Unknown
Trihydroxyoctadecadienoic acid
6′,7′Dihydroxybergamotin
Trihydroxyoctadecenoic acid
Unknown
Limonin/17Epilimonin.
Hydroxy-oxooctadecatrienoic acid
Hydroxy-oxooctadecenoic acid
Unknown
Hydroxy-oxooctadecadienoic acid
Name
+
-
-
Phenolic acid
+
-
-
Limonoid
-
-
+
Phenolic acid
+
-
-
Furanocoumarin
Prenylated sugar
O-flavanone
glycoside
-
-
+
+
-
-
+
-
+
+
-
Fatty acid
+
-
-
Furanocoumarin
-
-
+
Fatty acid
+
-
-
-
+
-
Limonoid
-
-
+
Fatty acid
+
-
-
Fatty acid
+
-
-
-
+
-
Fatty acid
+
-
-
Class
Red
cabbage
pomegran
ate
Grapefruit
)
70.
N.D.
586.3
71.
313.2373
C18H33O4
3.6
-
291.1966
C18H27O3
0.1
-
293.212
C18H29O3
0.8
275, 171
291.1967
C18H27O3
0.5
273
295.228
297.1519
C18H31O3
C19H21O3
0.4
-7.7
277
265
297.1527
297.1522
311.1678
C12H25O8
C19H21O3
C20H23O3
9.5
8.6
8
279, 175
-
277.2167
255.232
281.2485
339.1993
C18H29O2
C16H31O2
C18H33O2
C22H27O3
2.2
3.5
0.5
8.1
251, 211
-
N.D.
618.7
72.
N.D.
645.3
73.
673.3
74.
75.
76.
77.
78.
79.
80.
81.
82.
682.1
711.9
713.2
727.5
782.1
823.2
916.3
931.8
1008.9
278
N.D.
N.D.
N.D.
N.D.
N.D.
N.D.
N.D.
N.D.
N.D.
Dihydroxyoctadecenoic acid
Hydroxyoctadecatetraenoic acid
Hydroxyoctadecatrienoic acid
Oxo-octadecatrienoic
acid
Hydroxyoctadecadienoic acid
Geranyloxycoumarin
Hydroxy-octadecenoic
acid
Unknown
Unknown
Linolenic acid
Palmitic acid
Oleic acid
Unknown
Fatty acid
+
-
-
Fatty acid
+
-
-
Fatty acid
+
-
-
Fatty acid
+
-
-
Fatty acid
+
-
-
Coumarin
-
-
+
Fatty acid
+
-
-
+
+
+
-
+
+
+
+
-
Fatty acid
Fatty acid
Fatty acid
+ and – denotes the presence or absence of a metabolite in a certain functional food juice extract. N.D.= not detected